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Journal of Clinical Microbiology, September 2005, p. 4441-4447, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4441-4447.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development and Validation of an Immunoassay for Identification of Recent Human Immunodeficiency Virus Type 1 Infections and Its Use on Dried Serum Spots

Francis Barin,1* Laurence Meyer,2 Rémi Lancar,3 Christiane Deveau,2 Myriam Gharib,2 Anne Laporte,4 Jean-Claude Desenclos,4 and Dominique Costagliola3

Université François-Rabelais, EA3856 and Centre National de Référence du VIH, Tours,1 INSERM U569, Epidemiology Department, Hôpital Bicêtre AP-HP, Le Kremlin-Bicêtre,2 INSERM U720, Université Pierre et Marie Curie, Paris,3 Institut de Veille Sanitaire, Saint-Maurice, France4

Received 19 March 2005/ Returned for modification 12 May 2005/ Accepted 28 May 2005

The objective was to develop and to validate an immunossay to identify recent human immunodeficiency virus type 1 (HIV-1) infections that can be used on dried serum spots (DSS). A single, indirect enzyme-linked immunosorbent assay was developed to quantify antibodies toward four HIV-1 antigens: consensus peptides of the immunodominant epitope of gp41 (IDE), consensus V3 peptides, recombinant integrase, and recombinant p24. The parameters of the logistic regression used to classify the samples were estimated on a training sample (210 serum samples) using resampling techniques to get stable estimates and then applied to a validation sample (761 serum samples). The IDE and V3 peptides were the best able to discriminate between the antibodies present in serum from recently (≤6 months) infected individuals and those with long-lasting infection. Combined quantification of antibody binding to these two synthetic antigens allowed us to identify recent infections with an area under the receiver operating characteristic curve of 0.949 and a sensitivity of 88.3%, with a specificity of 97.6% in patients with long-term infection (but not AIDS) and 86.0% in patients suffering from AIDS with a threshold of 0.50 in the validation sample. This simple immunoassay can be used to identify recently HIV-1-infected patients. Its performance is compatible with its use in population-based studies including DSS.


* Corresponding author. Mailing address: Laboratoire de Virologie, Centre National de Référence du VIH, Centre Hospitalier Universitaire Bretonneau, 37044 Tours Cedex, France. Phone: 33 2 47 47 80 58. Fax: 33 2 47473610. E-mail: fbarin{at}med.univ-tours.fr.


Journal of Clinical Microbiology, September 2005, p. 4441-4447, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4441-4447.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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